Fig 2: Steroid hormones and ERK1/2 changes in the hypophysis of female DAT-KO rats. (A) Mineralocorticoid receptor (MCr) levels were increased in both DAT+/- and DAT-/- female rats, while Glucocorticoid receptor (GCr) levels were increased only in DAT-/- animals. (C) DAT +/- rats showed increase ERK1/2 protein levels. (B,D) Immunoblots from representative rats in each group are shown (PND60). (E) DAT ablation causes alterations of circulating blood levels of corticosterone in basal conditions and after stress. Data were normalized to ß-tubulin or ß-actin, relative to control (DAT +/+). Data were expressed as mean ± SEM (n = 4–6). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. ##p < 0.01 DAT-/- vs DAT+/+ and DAT+/- in basal conditions. ‡p < 0.05 DAT+/+ stress vs DAT-/- stress. ^ indicates same-gel/genotype/condition non-contiguous bands.

Fig 3: Dexamethasone induces nuclear translocation of the glucocorticoid receptor that is associated with cardiomyocyte hypertrophy. A, Neonatal rat ventricular cardiomyocytes were treated with dexamethasone (100 nmol/L) or ethanol for increasing time periods of 1, 6, or 24 hours before fixing and staining for glucocorticoid receptor, a-actinin (myocyte specificity), and DAPI (nucleus). Scale bar 10 µmol/L. B, Western blot showing cytoplasmic and nuclear fractions of neonatal rat ventricular myocytes cultured and treated with dexamethasone for increasing time periods of 1, 6, and 24 hours. Blots probed for glucocorticoid receptor (Nr3c1, nuclear receptor subfamily 3 group C member 1), GAPDH, and histone 2b. C, Signal intensities of the cytoplasmic glucocorticoid receptor normalized to GAPDH and nuclear glucocorticoid receptor normalized to histone 2b are shown. ImageJ was used for quantification. D, Graph represents cardiomyocyte area as measured using ImageJ software, tabulated and graphed in Prism 7 (GraphPad, La Jolla, CA) to show change in cell size with dexamethasone treatments, as indicated. All experiments shown were repeated as independent triplicates, *P<0.05. Dex indicates dexamethasone; GR, glucocorticoid receptor; H2b, histone 2b.